Phage Lysogeny and Inhibition of Biofilm Formation

噬菌体溶原性和生物膜形成的抑制

• 批准号: 7194399
• 负责人: George A. O'Toole
• 金额: $ 19.99万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-19 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7194399
• 关键词: Acute; Adhesions; Affect; Antibiotic Resistance; Arginine; Bacteremia; Bacteria; Bacteriophages; Burn injury; Carbon; Cells; Chronic; Clinical; Communities; Condition; DNA Sequence; Data; Development; Disease; Ecology; Elements; Employee Strikes; Epithelial Cells; Event; Evolution; Genes; Genetic; Genome; Genus Mycobacterium; Glass; Glucose; Goals; Growth; Immunity; In Vitro; Individual; Infection; Laboratories; Life Style; Lung; Lysogeny; Microbe; Microbial Biofilms; Modeling; Nature; Organism Strains; Outcome; Pathogenesis; Pathway interactions; Patients; Pattern; Plastics; Polymerase Chain Reaction; Polystyrenes; Property; Pseudomonas aeruginosa; Range; Reporting; Role; Role playing therapy; Sequence Analysis; Source; Suppurative Otitis Media; Surface; Testing; Toxin; Type III Secretion System Pathway; Virulence; Virulence Factors; Work; base; casamino acids; cross immunity; genome sequencing; improved; in vivo; insight; organic acid; prevent; trait

DESCRIPTION (provided by applicant): We previously reported the isolation and characterization of a transducing phage of P. aeruginosa, designated DMS3, originally isolated from a clinical strain of this organism. Recent studies in our lab have uncovered a new role for this lysogenic bacteriophage - the ability to inhibit the formation of surface- attached communities (known as biofilms) under all in vitro conditions tested. DMS3, when lysogenizing the laboratory strain P. aeruginosa PA14 or several clinical strains, results in a lysogen unable to make a biofilm under every condition we have tested, including growth on a variety of carbon sources (glucose, casamino acids, LB, arginine, organic acids) and on a variety of surfaces (i.e., PVC & polystyrene plastic, glass), and does so without affecting bacterial growth, indicating that lysogeny by phage DMS3 interferes with a central pathway required for biofilm formation by P. aeruginosa. While most of our studies have involved phage DMS3, we also have isolated several additional phages from clinical isolates of P. aeruginosa that also inhibit biofilm formation by P. aeruginosa. Preliminary evidence suggests that one such phage, designated DMS577, is distinct from DMS3. Taken together, these data indicate that phage DMS3 and other phages can modulate the ability of P. aeruginosa to initiate a surface-attached lifestyle. We hypothesize that lysogeny by phage DMS3 or DMS577 blocks biofilm formation by interfering with an essential component of the biofilm formation pathway. The interaction of bacteria and their phages are striking examples of polymicrobial infections resulting in the bacterium acquiring new virulence factors, evolving new pathogenic traits and increasing the ability of the phage-lysogenized microbe to cause disease in a wide range of hosts. Our data suggest that as part of a polymicrobial infection with P. aeruginosa, bacteriophages DMS3 and DMS577 may have the ability to influence the nature and outcome of infections, including chronic suppurative otitis media. To test the hypotheses above, we propose the following Specific Aims for this R21 application: Specific Aim I. Determine the phage DMS3-encoded functions required to inhibit biofilm formation. Specific Aim 2. Determine the step in biofilm formation blocked by phage lysogeny. Specific Aim III. Determine DMA sequence of phage DMS577 and the mechanism by which DMS577 interferes with biofilm formation.

描述（由申请人提供）：我们先前报道了铜绿假单胞菌转导噬菌体的分离和表征，命名为DMS 3，最初分离自该生物体的临床菌株。我们实验室最近的研究揭示了这种溶原性噬菌体的一个新作用-在所有体外测试条件下抑制表面附着群落（称为生物膜）形成的能力。DMS 3，当溶源化实验室菌株铜绿假单胞菌PA 14或几种临床菌株时，导致溶原菌在我们测试的每种条件下都不能形成生物膜，包括在各种碳源（葡萄糖、酪蛋白氨基酸、LB、精氨酸、有机酸）和各种表面（即，PVC和聚苯乙烯塑料，玻璃），并且这样做不影响细菌生长，表明噬菌体DMS 3的溶原性干扰铜绿假单胞菌形成生物膜所需的中心途径。虽然我们的大多数研究都涉及噬菌体DMS 3，但我们也从铜绿假单胞菌的临床分离株中分离出了几种额外的抑制铜绿假单胞菌生物膜形成的细菌。初步证据表明，一个这样的噬菌体，命名为DMS 577，是不同于DMS 3。综上所述，这些数据表明，噬菌体DMS 3和其他噬菌体可以调节铜绿假单胞菌启动表面附着生活方式的能力。 我们假设噬菌体DMS 3或DMS 577的溶原性通过干扰生物膜形成途径的重要组分来阻止生物膜形成。细菌和它们的病原体的相互作用是多微生物感染的突出例子，导致细菌获得新的毒力因子，进化新的致病性状，并增加噬菌体溶源化微生物在广泛宿主中引起疾病的能力。我们的数据表明，作为铜绿假单胞菌的多微生物感染的一部分，噬菌体DMS 3和DMS 577可能具有影响感染的性质和结果的能力，包括慢性化脓性中耳炎。 为了检验上述假设，我们为R21的应用提出了以下具体目标：具体目标I。确定抑制生物膜形成所需的噬菌体DMS 3编码功能。具体目标2。确定噬菌体溶原性阻断生物膜形成的步骤。具体目标三。确定噬菌体DMS 577的DMA序列和DMS 577干扰生物膜形成的机制。